Analysis of molecular domains of epitope-tagged merlin isoforms in Cos-7 cells and primary rat Schwann cells.
The Neurofibromatosis 2 gene product, merlin, has striking similarity to ezrin, radixin, and moesin (ERM), members of the protein 4.1 family which have been demonstrated to connect proteins in the plasma membrane to the cytoskeletal components. The recent localization of merlin to the motile regions in cultured cells such as membrane ruffles further supports the notion that merlin represents a new class of tumor suppressors. Here we describe the localization of full-length and truncated polypeptides of merlin expressed as Flag-tagged proteins in transfected cells. Similar to endogenous merlin, the epitope-tagged full-length merlin localizes to the membrane ruffles in transfected Cos-7 cells and rat Schwann cells. In addition, the over-expressed merlin localizes to other actin-rich cortical structures, such as microvilli and filopodia. The amino-terminal half of merlin is seen dispersed throughout the cells and in membrane ruffles. Compared to the amino-terminal half of merlin, its carboxy-terminal half localizes more distinctly to membrane ruffles. The full-length and the carboxy-terminal portion of merlin co-localize with F-actin at the membrane ruffles. However, distinct from the ERM proteins, the carboxy-terminal-truncated merlin and F-actin do not co-localize with each other at the stress fibers. Our results suggest that both the amino- and the carboxy-terminal domains of merlin contribute to its membrane ruffle localization.
Xu
L
L
Molecular Neurogenetics Unit, Massachusetts General Hospital East, Charlestown 02129, USA.
Gonzalez-Agosti
C
C
Beauchamp
R
R
Pinney
D
D
Sterner
C
C
Ramesh
V
V
D000818:Animals; D000831:Animals, Newborn; D019556:COS Cells; D002455:Cell Division; D002478:Cells, Cultured; D016515:Genes, Neurofibromatosis 2; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D025581:Neurofibromin 2; D009842:Oligopeptides; D010455:Peptides; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis; D012583:Schwann Cells / Q000166:cytology / Q000378:metabolism*; D016324:Sequence Tagged Sites; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D014162:Transfection
D008565:Membrane Proteins; D025581:Neurofibromin 2; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins; C085706:FLAG peptide